家兔产气荚膜梭菌的分离、鉴定及致病性分析

Isolationand pathogenicity analysis of Clostridium perfringens in rabbits

为对疑似家兔产气荚膜梭菌病感染死亡的肉兔病例进行病原菌的分离鉴定,并分析其对家兔的致病性,试验采集死亡病例的肠道内容物,划线接种于兔鲜血平板和TSC选择鉴别培养基中,对病原菌进行分离纯化,利用细菌16SrDNA通用引物进行分子鉴定,分析其同源性,构建系统进化发育树,同时,对病原菌进行生化鉴定,并将病原菌腹腔接种30日龄肉兔,分析其致病性。结果表明,从病料中分离纯化到1株革兰氏阳性菌;通过对细菌16S rDNA基因测序分析及生化鉴定确定为产气荚膜梭菌,且在系统进化树中也与其聚为一簇;致病性分析发现,该菌对家兔具有较强的致死性,可使试验组肉兔在攻毒16h后全部死亡。综上所述,本研究成功获得1株具有较强毒性的兔源产气荚膜梭菌,为产气荚膜梭菌致病机制的研究奠定了基础,同时也为疫病诊断和防控提供了理论依据。

In order to isolate pathogenic bacteria of the rabbit with suspected Clostridium perfringens infection,and analyze the pathogenicity in rabbits.The intestinal contents of death cases were collected and scored.Inoculated on rabbit blood plate and TSC selection identification medium,the pathogens were isolated and purified,and the 16S rDNA universal primers were used for molecular identification,and the homology was analyzed to construct a phylogenetic tree.At the same time,the pathogens were biochemically identified.The pathogenic bacteria were inoculated intraperitoneally into 30-day-old meat rabbits to analyze their pathogenicity.The results showed that a strain of Gram-positive bacteria was isolated and purified from the disease material;it was identified as Clostridium perfringens by sequencing and biochemical identification of the 16S rDNA gene,and it was also clustered with Clostridium perfringens in the phylogenetic tree.Pathogenicity analysis found that the strain had strong lethality to rabbits,and it could make the test group meat rabbits all died after 16h of attack.In summary,this study successfully obtained a highly virulent rabbit-producing Clostridium perfringens,which laid the foundation for the research on the pathogenic mechanism of Clostridium perfringens,and provided a theoretical basis for the diagnosis and prevention of the disease.